- Home
- IVD
- By Technology Types
- By Diseases Types
- By Product Types
- Research
- Resource
- Distributors
- Company
| Product Name | III One-Step RT-qPCR Probe Kit |
| Catalog No. | RESK-0068 |
| Description | The III One-Step RT-qPCR Probe Kit is designed for quantitative PCR using RNA templates. Reverse transcription and quantitative PCR proceed in the same reaction tube, simplifying handling and reducing contamination risk. Using gene-specific primers, a thermostable reverse transcriptase efficiently synthesizes first-strand cDNA, which is then quantified with a fluorescence-labeled probe and an antibody-based hot-start Taq DNA polymerase. Paired with an optimized buffer system, the kit enables specific and sensitive quantification of RNA templates. The kit is supplied as a convenient premix: the 2x P buffer contains the optimized buffer system and dNTPs, optimized with factors that effectively suppress nonspecific PCR amplification and factors that enhance amplification efficiency, and is suited to high-specificity detection systems based on fluorescence-labeled probes. The UH III Enzymes blend contains an optimally proportioned mix of reverse transcriptase, RNase inhibitor, and hot-start Taq DNA polymerase. |
| Intended Use | Probe-based one-step RT-qPCR for specific and sensitive quantification of RNA templates. |
| Principle / Technology | One-tube reverse transcription and probe-based qPCR with gene-specific primers; thermostable reverse transcriptase for first-strand cDNA synthesis; antibody-based hot-start Taq DNA polymerase; buffer optimized with nonspecific-amplification inhibitors and amplification-enhancing factors. |
| Detection Method | Fluorescence-labeled probe (TaqMan probe) based detection. |
| Sample Type | RNA templates. |
| Performance Range / Specifications | High sensitivity. Annealing temperature 60°C; GC range 30%-70%; amplicon length 80-300 bp. |
| Sensitivity / LOD | High sensitivity detection of RNA templates. |
| Specificity | Probe-based high-specificity detection; buffer components effectively suppress nonspecific PCR amplification. |
| Reaction Conditions / Protocol | Reverse transcription and qPCR performed sequentially in the same tube; recommended annealing temperature 60°C; amplicon length 80-300 bp. For three-step cycling programs, set signal acquisition at the 72°C extension step; for two-step programs, acquire signal during the annealing/extension step. A cycle number of 40 is generally recommended. |
| Components / Formulation | 2x III P Buffer: 500 μL (50 T) / 2x1 mL (200 T) UH III Enzymes: 50 μL (50 T) / 200 μL (200 T) 50x High ROX: 20 μL (50 T) / 80 μL (200 T) 50x Low ROX: 20 μL (50 T) / 80 μL (200 T) RNase-free ddH2O: 500 μL (50 T) / 2x1 mL (200 T) |
| Storage Conditions | Store at -20°C, protected from light. Avoid repeated freeze-thaw cycles; aliquoting is recommended. |
| Shelf Life | 1 year when stored at -20°C, protected from light. |
| Package Specifications | 50 T; 200 T |
| Product Form | Mix |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | 1 year at -20°C protected from light. |
| Compatibility | Compatible with the vast majority of real-time PCR instruments; ROX reference dye (50x High ROX and 50x Low ROX) is supplied separately and added as required by the instrument. |
| Recommended Buffer System | 2x optimized probe buffer containing dNTPs, nonspecific-amplification inhibitors, and amplification-enhancing factors (included). |
| Application Notes / Precautions | Poor standard curve (amplification efficiency <90% or >110%): increase the dilution factor and pipetting volume for greater accuracy; re-prepare degraded standards; dilute overly concentrated templates that may inhibit the reaction; redesign primers with poor specificity. No amplification curve: increase cycle numbers appropriately (40 is typical; too many cycles raise background); confirm a signal acquisition step is set (72°C extension for three-step programs; annealing/extension for two-step programs); verify primer integrity by PAGE if primers have been stored long; reduce dilution if template concentration is too low; re-prepare degraded templates. Poor reproducibility: calibrate pipettes regularly and add templates at high dilution in larger volumes; calibrate the instrument for positional temperature consistency; note that very low template concentrations reduce reproducibility; run 4-6 replicates and exclude inconsistent wells. Template diluent: Nuclease-Free water, DEPC-treated water, or laboratory ddH2O are all acceptable. TE is not recommended because EDTA inhibits enzyme activity. |
For research use only, not for clinical use.
|
There is no product in your cart. |
Copyright © 2026 Alta DiagnoTech. All rights reserved.
